Protein Purification from Corn Germ .


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Protein Decontamination from Corn Germ. Danielle McConnell Division of Concoction Building Iowa State College. Why separate Protein X from Transgenic Corn?. Could be a decent option for a low calorie sweetener 9500 times sweeter than sucrose on a sub-atomic premise
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Protein Purification from Corn Germ Danielle McConnell Department of Chemical Engineering Iowa State University

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Why separate Protein X from Transgenic Corn? Could be a decent option for a low calorie sweetener 9500 circumstances sweeter than sucrose on an atomic premise Good extraction properties (warm steady and low sub-atomic weight) Transgenic corn is financially plausible, simple to scale up, and is very accessible

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Extraction (section 1) pH assurance Isoelectric purpose of Protein is 5.72 Tested pHs 4,5,6, and 7 Total Protein Assays pH 4 demonstrated minimal measure of aggregate protein separated

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Extraction (section 2) Determination of Heating Time At 80 ° C Protein is thermo-stable Total Protein Assays of accelerate at various circumstances At ten minutes the aggregate protein is the littlest

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Cation Exchange Chromatography Proteins tie to the contrarily charged groups by electrostatic powers A SP-Sepharose segment was utilized to run the particle trade According to the chromatogram (see next board), top 6 or division #38 shows the protein rich example Cation Exchange Setup appeared above

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Cation Exchange Chromatogram Conductivity versus Time (part # at top of diagram)

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Ultrafiltration Used an utilized extensive and little atomic weight cutoff layers to channel the concentrate Protein ought to be left in the retentate of the little MW cutoff film Total Protein Assays gave a preparatory assessment of what was left in the retentate

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Ultrafiltration Flowchart

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Detects virtue of Protein from infused test Concentration can be found by incorporating the pinnacle range Protein X ordinarily elutes around 15 minutes The HPLC chromatogram shows three elutions. The red line demonstrates an immaculate protein test, the blue line speaks to unadulterated protein from the cation trade, and the dark line depicts a specimen of concentrate with spiked protein . Turn around Phase HPLC

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HPLC Chromatogram mV versus Minutes

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Final Discussion: The last consequences of this venture have not been resolved, be that as it may, numerous dynamic strides have been made in getting the protein from corn germ. Different extraction conditions (pH and salt conditions), filtration steps, and investigation of an evaluation strategy (ELISA tests) are at present being looked into to acquire a cleansed protein. The last strides of the protein grouping will be founded on a mix of elements, for example, virtue and yield which will be tried by switch stage HPLC and the ELISA test. Graph of finish Antibody setup for ELISA testing

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Acknowledgments: A developed satisfaction and thank you is made to the large number of individuals who made this venture conceivable as well as fruitful: Dr. Charles Glatz PWSE Yandi Dharmadi, Zhengrong Gu, Maureen Griffin, Todd Menkhaus, and Jim Kupferschmit

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